• 한국토양비료학회지
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• 제49권4호
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• pp.393-400
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• 2016

Fusarium graminearum is the main cause of substantial economic loss in wheat production. The aim of this study is to investigate biocontrol potential of Lysobacter antibioticus HS124 against F. graminearum and to purify an antifungal compound. In preliminary study, n-butanol crude extract revealed destructive alterations in the hyphal morphology of F. graminearum and almost degraded with $1,000{\mu}g\;mL^{-1}$ concentration. For further study, the antifungal compound extracted from the n-butanol crude extract of L. antibioticus HS124 was identified as N-Butyl-tetrahydro-5-oxofuran-2-carboxamide ($C_9H_{16}NO_3$) using NMR ($^1H-NMR$, $^{13}C-NMR$, $^1H-^1H\;COSY$, HMBC, and HMQC), and HR-ESI-MS analysis. To our knowledge, N-Butyl-tetrahydro-5-oxofuran-2-carboxamide may be a novel compound with molecular weight of 186.1130. The minimum inhibitory concentration value of antifungal compound was $62.5{\mu}g\;mL^{-1}$ against F. graminearum. In an in vivo pot experiment, crown rot disease from F. graminearum was inhibited when wheat seeds were treated with both HS124 culture and F. graminearum. Growth of wheat seedling was enhanced by treatment of HS124 compared to control. Our results suggest that L. antibioticus HS124 characterized in this study could be successfully used to control F. graminearum and could be used as an alternative to chemical fungicides in modern agriculture.

• 미생물학회지
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• 제31권4호
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• pp.312-317
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• 1993

강원도와 남부지방에서 각각 옥수수와 보리를 채집하여 두 곡류로 부터 총 110 균주의 Fusatium graminearum을 분리한 후 쌀배지에서의 trichothecene과 zearalenone(ZEA)의 생성능을 측정하였다. 옥수수로 부터 분리한 51개의 F. graminearum 균주중 trichothecene 생성빈도는 deoxynivalenol(Don)이 64.5%, 3-acetyldeoxynivalenol (3-ADON)이 7.8%, 15acetyldeoxynivalenol (15-ADON)이 33.3%, nivalenol(NIV)이 21.6%, 4-acetylnivalenol (4-ANIV)이 13.7%였다. DON을 생성하는 균주들은 3-ADON보다는 15-ADON을 동시에 생성하였다. 한편, 보리로부터 분리한 59개의 F. graminearum 균주중 trichothecene 생성빈도는 NIV가 71.2%, 그리고 4-ANIV가 61.0%였으며, 한 균주만이 DON과 3-ADON을 동시에 생성하였다. 옥수수로부터 분리한 균주들의 ZEA의 생성빈도와 생성량은 각각 32.0%, 71㎍/g였으며, 보리로부터 분리한 균주는 각각 29%, 745㎍이었다. 이와같이 우리나라 강원도지방과 남부지방의 F. graminearum 균주들의 trichothecene 생성양상은 지역적인 큰 차이를 나타내었다.

• 식물병연구
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• 제16권3호
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• pp.306-311
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• 2010

2009년 국내산 옥수수 19점과 벼 32점을 대상으로 Fusarium 오염 및 Fusarium 독소 오염을 조사하였다. 옥수수와 벼 시료의 Fusarium 오염률은 각각 54.9%와 8.2%로 확인되었으며, 종 특이 primer를 이용한 PCR 증폭결과 옥수수시료에서 분리된 506균주 중 58균주의 F.graminearum 추정균주(11.5%)와 354균주의 F. verticillioides 추정균주(70.0%)가 동정되었다. 또한 벼의 경우, 분리된 315균주 중 276균주(87.8%)가 F. graminearum으로 추정 되었으며, F. verticillioides 추정균주는 검출되지 않았다. LC 및 LC-MS를 이용한 Fusarium 독소(DON, NIV, ZEA, FB)의 자연발생량 조사 결과, DON과 ZEA이 각각 2개의 옥수수 시료에서만 기준치 이상 검출되었다. FB는 대부분의 옥수수 시료와 한 개의 벼 시료에서 검출 되었으나 모두 기준치 이하였다. 따라서 본 연구에서 사용된 2009년산 옥수수와 벼의 곰팡이독소 오염수준은 대부분 기준치 이하로 심각하지 않았다.

• 한국균학회지
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• 제37권2호
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• pp.121-129
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• 2009

In 2007, a total of 77 isolates of Fusarium spp. were obtained from ear rot symptoms of corns collected from 5 locations in Gangwon Province, Korea. The fungal isolates were identified based on their morphological features. Out of the isolates, fifteen isolates were identified as Fusarium verticillioides which formed microconidia in long chains on monophialides. Four isolates were identified as F. subglutinans which formed microconida only on false heads. Six isolates were identified as F. graminearum which produced red pigment in PDA culture. Besides these Fusarium species, F. napiform, F. nygamai, and F. oxysporum were identified from the rest isolates. To assess for genetic diversity of the isolates, a random amplified polymorphic DNA(RAPD) technique was carried out using URP primers. The results from the RAPD analysis showed that the isolates from corn were divided into 6 groups. These RAPD groups of the Fusarium species corresponded to morphological characters of the Fusarium species. The phylogenetic analysis of most isolates by DNA sequencing of EF-1$\alpha$ gene corresponded to morphological characters of the Fusarium species. The results of pathogenicity tests by two inoculation methods revealed that F. verticillioides, F. graminearum and F. subglutinans are strongly pathogenic to corn stalks.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 추계학술대회 및 정기총회
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• pp.15-15
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• 2014

Fusarium graminearum (teleomorph Gibberella zeae) is an important plant pathogen that causes head blight of major cereal crops such as wheat, barley, and rice, as well as causing ear and stalk rot on maize worldwide. Plant diseases caused by this fungus lead to severe yield losses and accumulation of harmful mycotoxins in infected cereals [1]. Fungi utilize spore production as a mean to rapidly avoid unfavorable environmental conditions and to amplify their population. Spores are produced sexually and asexually and their production is precisely controlled. Upstream developmental activators consist of fluffy genes have been known to orchestrate early induction of condiogenesis in a model filamentous fungus Aspergillus nidulans. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we characterized functions of the F. graminearum fluffy gene homologs [2]. We found that FlbD is conserved regulatory function for conidiogenesis in both A. nidulans and F. graminearum among five fluffy gene homologs. flbD deletion abolished conidia and perithecia production, suggesting that FlbD have global roles in hyphal differentiation processes in F. graminearum. We further identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum [3]. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. F. graminearum ortholog of Aspergillus nidulans wetA has been shown to be involved in conidiogenesis and conidium maturation [4]. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidia dormancy by suppressing microcycle conidiation in F. graminearum. In A. nidulans, FlbB physically interacts with FlbD and FlbE, and the resulting FlbB/FlbE and FlbB/FlbD complexes induce the expression of flbD and brlA, respectively. BrlA is an activator of the AbaA-WetA pathway. AbaA and WetA are required for phialide formation and conidia maturation, respectively [5]. In F. graminearum, the AbaA-WetA pathway is similar to that of A. nidulans, except a brlA ortholog does not exist. Amongst the fluffy genes, only fgflbD has a conserved role for regulation of the AbaA-WetA pathway.

• 한국균학회지
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• 제13권2호
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• pp.99-103
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• 1985

품종(品種)별로 포장(圃場)에서 임의(任意)로 채집(採集)하여 병원균(病原菌)을 분리(分離)한 결과(結果) Fusarium moniliforme와 F. roseum 'Graminearum' 품종(品種)에 따라 분리(分離) 비율(比率)이 다르나 공시(供試)한 품종(品種)에서 거의 다 분리(分離)되었다. 특(特)히 F. roseum 'Graminearum'은 현미(玄米)에서도 높은 분리비율(分離比率)을 보였다. 참새 피해(피해)를 받은 이삭에서 F. moniliforme의 Sporodochia가 많이 형성(形成)되었으며 벼 별구에서도 . F. moniliforme등 많은 병원균(病原菌)이 분리(分離)되었다. 저지원 시험결과 P242가 F. moniliforme에 대하여 저지비교(效果)가 가장 좋았으나 소독(消毒)후 종피(種皮)에서는 병원균(病原菌)이 거의 분리(分離)되지 않았으나 현미(玄米)에서는 키다리병원균(病原菌)인 F. moniliforme가 많이 분리(分離)되었다. 소독시간별(消毒時間別) 약효(藥效)를 보면 4시간 동안 처리하면 12시간 24시간 36시간 보다 약효가 띨어졌으며 $30^{\circ}C$에서 36 시간 처리한것이 가장 종은 것으로 나타났다.

• The Plant Pathology Journal
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• 제15권1호
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• pp.53-56
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• 1999

Fifty-three isolates of Fusarium graminearum were obtained from corn and barley samples in several provinces of Korea. Gas chromatography-mass spectrometric analysis of trichothecenes produced by these isolates revealed that 37 and 16 isolates were nivalenol (NIV)- and deoxynivalenol (DON)-chemotypes, respectively. Two hundred and seventy-five nitrate-nonutilizing (nit) mutants were obtained from the isolates. Of these mutants, 187 were identified as nit1, nit3, or NitM, but 88 could not be identified as one of these classes. The highest frequency of nit mutant was nit1 (65%), followed by nit3 (20%) and NitM (15%). Higher frequency of NitM was observed in DON-chemotypes than in NIV-chemotypes. The mutants were used for vegetative compatibility group (VCG) analysis by examining heterokaryosis using complementary mutant pairs. No heterokaryon formation was observed among all 1,248 pairwise combinations, suggesting that all isolates tested belong to different VCGs. Higher frequency of self-incompatibility was observed in NIV-chemotypes than in DON-chemotypes. These results suggest that the like-lihood of asexual genetic recombination may be very low I F. graminearum under the field condition.

• The Plant Pathology Journal
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• 제27권4호
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• pp.301-309
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• 2011

The filamentous fungus Fusarium graminearum is an important cereal pathogen. Although quantitative realtime PCR (qRT-PCR) is commonly used to analyze the expression of important fungal genes, no detailed validation of reference genes for the normalization of qRT-PCR data has been performed in this fungus. Here, we evaluated 15 candidate genes as references, including those previously described as housekeeping genes and those selected from the whole transcriptome sequencing data. By a combination of three statistical algorithms (BestKeeper, geNorm, and NormFinder), the variation in the expression of these genes was assessed under different culture conditions that favored mycelial growth, sexual development, and trichothecene mycotoxin production. When favoring mycelial growth, GzFLO and GzUBH expression were most stable in complete medium. Both EF1A and GzRPS16 expression were relatively stable under all conditions on carrot agar, including mycelial growth and the subsequent perithecial induction stage. These two genes were also most stable during trichothecene production. For the combined data set, GzUBH and EF1A were selected as the most stable. Thus, these genes are suitable reference genes for accurate normalization of qRT-PCR data for gene expression analyses of F. graminearum and other related fungi.

• The Plant Pathology Journal
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• 제30권1호
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• pp.33-42
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• 2014

Nivalenol (NIV) and deoxynivalenol (DON) are predominant Fusarium-producing mycotoxins found in grains, which are mainly produced by Fusarium asiaticum and F. graminearum. NIV is found in most of cereals grown in Korea, but the genetic basis for NIV production by F. asiaticum has not been extensively explored. In this study, 12 genes belonging to the trichothecene biosynthetic gene cluster were compared at the transcriptional level between two NIV-producing F. asiaticum and four DON-producing F. graminearum strains. Chemical analysis revealed that time-course toxin production patterns over 14 days did not differ between NIV and DON strains, excluding F. asiaticum R308, which was a low NIV producer. Both quantitative real-time polymerase chain reaction and Northern analysis revealed that the majority of TRI gene transcripts peaked at day 2 in both NIV and DON producers, which is 2 days earlier than trichothecene accumulation in liquid medium. Comparison of the gene expression profiles identified an NIV-specific pattern in two transcription factor-encoding TRI genes (TRI6 and TRI10) and TRI101, which showed two gene expression peaks during both the early and late incubation periods. In addition, the amount of trichothecenes produced by both DON and NIV producers were correlated with the expression levels of TRI genes, regardless of the trichothecene chemotypes. Therefore, the reduced production of NIV by R308 compared to NIV or DON by the other strains may be attributable to the significantly lower expression levels of the TRI genes, which showed early expression patterns.

• The Plant Pathology Journal
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• 제30권1호
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• pp.25-32
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• 2014

Fusarium head blight (FHB; scab) caused mainly by Fusarium graminearum is a devastating disease of wheat and barley around the world. FHB causes yield reductions and contamination of grain with trichothecene mycotoxins such as deoxynivalenol (DON) which are a major health concern for humans and animals. The objective of this research was to develop an easy seed or seedling inoculation assay, and to compare these assays with whole plant resistance of twenty-nine Korean winter wheat cultivars to FHB. The clip-dipping assay consists of cutting off the coleoptiles apex, dipping the coleoptiles apex in conidial suspension, covering in plastic bag for 3 days, and measuring the lengths of lesions 7 days after inoculation. There were significant cultivar differences after inoculation with F. graminearum in seedling relative to the controls. Correlation coefficients between the lesion lengths of clip-dipping inoculation and FHB Type II resistance from adult plants were significant (r=0.45; P<0.05). Results from two other seedling inoculation methods, spraying and pin-point inoculation, were not correlated with adult FHB resistance. Single linear correlation was not significant between seed germination assays (soaking and soak-dry) and FHB resistance (Type I and Type II), respectively. These results showed that clip-dipping inoculation method using F. graminearum may offer a real possibility of simple, rapid, and reliable for the early screening of FHB resistance in wheat.